LOCUS       sequence-242695-        8769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Myc-tagged LATS2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  AUTHORS   Bao Y, Nakagawa K, Yang Z, Ikeda M, Withanage K, Ishigami-Yuasa M, 
            Okuno Y, Hata S, Nishina H, Hata Y
  TITLE     A cell-based assay to screen stimulators of the Hippo pathway 
            reveals the inhibitory effect of dobutamine on the YAP-dependent 
            gene transcription.
  JOURNAL   J Biochem. 2011 Aug;150(2):199-208. doi: 10.1093/jb/mvr063. Epub 
            2011 May 17.
  PUBMED    21586534
REFERENCE   2  (bases 1 to 8769)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1093/jb/mvr063";
            journalName: "J Biochem"; date: "2011-08"; volume: "150"; issue: 
            "2"; pages: "199-208"
COMMENT     Sequence Label: pClneoMyc-LATS2
FEATURES             Location/Qualifiers
     source          1..8769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     708..732
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     987..1006
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1049..1067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1085..1114
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4455..4576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4466..4485)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4520..4539
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4762..5217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5334..5691
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5334..5525
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5357..5377)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5542..5677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5742..6536
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5796..5815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6406..6425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6600..6648
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6689..6708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6808..6830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6868..6886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6954..7058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7059..7127,7128..7919)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7277..7296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8090..8678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8579..8598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catattattc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
       61 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
      121 gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tgcgatcgcc cgccccgttg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      721 aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat tgctaacgca
      781 gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gtgactctct taaggtagcc
      841 ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
     1021 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gaccccccaa
     1081 catggagcag aagcttatca gcgaggagga cctgctcgag gaattcacgc gtatgaggcc
     1141 aaagactttt cctgccacga cttattctgg aaatagccgg cagcgactgc aagagattcg
     1201 tgaggggtta aagcagccat ccaagtcttc ggttcagggg ctacccgcag gaccaaacag
     1261 tgacacttcc ctggatgcca aagtcctggg gagcaaagat gccaccaggc agcagcagca
     1321 gatgagagcc accccaaagt tcggacctta tcagaaagcc ttgagggaaa tcagatattc
     1381 cttgttgcct tttgctaatg aatcgggcac ctctgcagct gcagaagtga accggcaaat
     1441 gctgcaggaa ctggtgaacg caggatgcga ccaggagatg gctggccgag ctctcaagca
     1501 gactggcagc aggagcatcg aggccgccct ggagtacatc agcaagatgg gctacctgga
     1561 cccgaggaat gagcagattg tgcgggtcat taagcagacc tccccaggaa aggggctcat
     1621 gccaacccca gtgacgcgga ggcccagctt cgaaggaacc ggcgattcgt ttgcgtccta
     1681 ccaccagctg agcggtaccc cctacgaggg cccaagcttc ggcgctgacg gccccacggc
     1741 gctggaggag atgccgcggc cgtacgtgga ctaccttttc cccggagtcg gcccccacgg
     1801 gcccggccac cagcaccagc acccacccaa gggctacggt gccagcgtag aggcagcagg
     1861 ggcacacttc ccgctgcagg gcgcgcacta cgggcggccg cacctgctgg tgcctgggga
     1921 acccctgggc tacggagtgc agcgcagccc ctccttccag agcaagacgc cgccggagac
     1981 cgggggttac gccagcctgc ccacgaaggg ccagggagga ccgccaggcg ccggcctcgc
     2041 tttcccaccc cctgccgccg ggctctacgt gccgcaccca caccacaagc aggccggtcc
     2101 cgcggcccac cagctgcatg tgctgggctc ccgcagccag gtgttcgcca gcgacagccc
     2161 cccgcagagc ctgctcactc cctcgcggaa cagcctcaac gtggacctgt atgaattggg
     2221 cagcacctcc gtccagcagt ggccggctgc caccctggcc cgccgggact ccctgcagaa
     2281 gccgggcctg gaggcgccgc cgcgcgcgca cgtggccttc cggcctgact gcccagtgcc
     2341 cagcaggacc aactccttca acagccacca gccgcggccc ggtccgcctg gcaaggccga
     2401 gccctccctg cccgccccca acaccgtgac ggctgtcacg gccgcgcaca tcttgcaccc
     2461 ggtgaagagc gtgcgtgtgc tgaggccgga gccgcagacg gctgtggggc cctcgcaccc
     2521 cgcctgggtg cccgcgcctg ccccggcccc cgcccccgcc cccgccccgg ctgcggaggg
     2581 cttggacgcc aaggaggagc atgccctggc gctgggcggc gcaggcgcct tcccgctgga
     2641 cgtggagtac ggaggcccag accggaggtg cccgcctccg ccctacccga agcacctgct
     2701 gctgcgcagc aagtcggagc agtacgacct ggacagcctg tgcgcaggca tggagcagag
     2761 cctccgtgcg ggccccaacg agcccgaggg cggcgacaag agccgcaaaa gcgccaaggg
     2821 ggacaaaggc ggaaaggata aaaagcagat tcagacctct cccgttcccg tccgcaaaaa
     2881 cagcagagac gaagagaaga gagagtcacg catcaagagc tactcgccat acgcctttaa
     2941 gttcttcatg gagcagcacg tggagaatgt catcaaaacc taccagcaga aggttaaccg
     3001 gaggctgcag ctggagcaag aaatggccaa agctggactc tgtgaagctg agcaggagca
     3061 gatgcggaag atcctctacc agaaagagtc taattacaac aggttaaaga gggccaagat
     3121 ggacaagtct atgtttgtca agatcaaaac cctggggatc ggtgcctttg gagaagtgtg
     3181 ccttgcttgt aaggtggaca ctcacgccct gtacgccatg aagaccctaa ggaaaaagga
     3241 tgtcctgaac cggaatcagg tggcccacgt caaggccgag agggacatcc tggccgaggc
     3301 agacaatgag tgggtggtca aactctacta ctccttccaa gacaaagaca gcctgtactt
     3361 tgtgatggac tacatccctg gtggggacat gatgagcctg ctgatccgga tggaggtctt
     3421 ccctgagcac ctggcccggt tctacatcgc agagctgact ttggccattg agagtgtcca
     3481 caagatgggc ttcatccacc gagacatcaa gcctgataac attttgatag atctggatgg
     3541 tcacattaaa ctcacagatt tcggcctctg cactgggttc aggtggactc acaattccaa
     3601 atattaccag aaagggagcc atgtcagaca ggacagcatg gagcccagcg acctctggga
     3661 tgatgtgtct aactgtcggt gtggggacag gctgaagacc ctagagcaga gggcgcggaa
     3721 gcagcaccag aggtgcctgg cacattcact ggtggggact ccaaactaca tcgcacccga
     3781 ggtgctcctc cgcaaagggt acactcaact ctgtgactgg tggagtgttg gagtgattct
     3841 cttcgagatg ctggtggggc agccgccctt tttggcacct actcccacag aaacccagct
     3901 gaaggtgatc aactgggaga acacgctcca cattccagcc caggtgaagc tgagccctga
     3961 ggccagggac ctcatcacca agctgtgctg ctccgcagac caccgcctgg ggcggaatgg
     4021 ggccgatgac ctgaaggccc accccttctt cagcgccatt gacttctcca gtgacatccg
     4081 gaagcagcca gccccctacg ttcccaccat cagccacccc atggacacct cgaatttcga
     4141 ccccgtagat gaagaaagcc cttggaacga tgccagcgaa ggtagcacca aggcctggga
     4201 cacactcacc tcgcccaata acaagcatcc tgagcacgca ttttacgaat tcaccttccg
     4261 aaggttcttt gatgacaatg gctacccctt tcgatgccca aagccttcag gagcagaagc
     4321 ttcacaggct gagagctcag atttagaaag ctctgatctg gtggatcaga ctgaaggctg
     4381 ccagcctgtg tacgtgtagt cgacccgggc ggccgcttcc ctttagtgag ggttaatgct
     4441 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4501 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     4561 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     4621 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg
     4681 atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4741 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     5221 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     5281 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     5341 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5461 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5521 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5581 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5641 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     5701 tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat
     5761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6241 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     6301 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6541 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     6601 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     6661 taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6721 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6781 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6841 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     6901 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6961 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7021 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7141 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     7201 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     7261 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     7321 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     7381 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     7441 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     7501 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7561 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7621 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7681 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7741 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7801 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7861 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7921 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7981 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     8161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8221 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     8521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8701 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc ttcaatattg
     8761 gccattagc
//
